*4.3. Preparation of TO@NZ Hybrid Nanostructures*

The preparation of TO@NZ bioactive hybrid nanostructures was carried out based on the method reported elsewhere for nanoclays modification [15] with some small changes. In a spherical flask, an amount of 20 mL TO were heated to boil. Before its use, the thyme oil was freed up from the Limonene and Cymene by distillation process at 190 ◦C. An amount of 6 mL was removed which is approximately 30% of the total amount. This result is in close agreement with the GC measurements data. The distillation process was the first stage of the overall process which was followed in this study. The distillation temperature was chosen a little higher than Limonene's and Cymene's boiling point temperatures (170–175 ◦C) and lower than thymol's boiling point temperature (230–235 ◦C). After the first stage, the remaining liquid colour was changed to dark brown and in line with LC-MS/MS measurements, it was rich of thymol oil. In the second step, an evaporation-adsorption process was carried out. The remained liquid from the first stage was boiled up to 250 ◦C. The produced thymol rich steam passed through a bed of 3 gr NZ which strongly adsorbed the essential oil. The remaining outlet steam was cooled down through a condenser and the final liquid was collected. At the same time, an amount of as received thyme oil was adsorbed by NZ material and reference hybrid nanostructured material TO\_NZ was prepared. The %wt. adsorbed TO in both TO@NZ and TO\_NZ hybrid nanostructures was calculated gravimetrically and was approx. 80% wt. for both hybrid nanostructures.
